A high-throughput screen identifies that CDK7 activates glucose consumption in lung cancer cells
A high-throughput screen identifies that CDK7 activates glucose consumption in lung cancer cells
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DOI：
10.1038/s41467-019-13334-8
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发表时间：
2019-11-29
影响因子：
16.6
通讯作者：
Clark, Peter M.
中科院分区：
文献类型：
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作者：
Ghezzi, Chiara;Wong, Alicia;Clark, Peter M.
葡萄糖消耗增加是癌症的基础，但选择性地针对这一途径是具有挑战性的。我们开发了一种用于测量葡萄糖消耗的高通量检测方法，并用它来筛选具有生物活性的小分子对非小细胞肺癌细胞系的作用。我们发现Milciclib可以通过降低SLC2A1(GLUT1)的mRNA和蛋白水平以及抑制葡萄糖运输来阻止H460和H1975的葡萄糖消耗，但在HCC827或A549细胞中没有。Milciclib通过靶向细胞周期蛋白依赖性激酶7(CDK7)来阻止葡萄糖消耗，类似于其他CDK7抑制剂，包括THZ1和LDC4297。增强的PIK3CA信号导致CDK7磷酸化，从而促进RNA聚合酶II的磷酸化和转录。Milciclib、THZ1和LDC4297导致SLC2A1启动子上的RNA聚合酶II磷酸化降低。这些数据表明，我们的高通量分析可以识别调节葡萄糖消耗的化合物，并且CDK7是具有激活的PI3K途径的细胞中葡萄糖消耗的关键调节因子。
Elevated glucose consumption is fundamental to cancer, but selectively targeting this pathway is challenging. We develop a high-throughput assay for measuring glucose consumption and use it to screen non-small-cell lung cancer cell lines against bioactive small molecules. We identify Milciclib that blocks glucose consumption in H460 and H1975, but not in HCC827 or A549 cells, by decreasing SLC2A1 (GLUT1) mRNA and protein levels and by inhibiting glucose transport. Milciclib blocks glucose consumption by targeting cyclin-dependent kinase 7 (CDK7) similar to other CDK7 inhibitors including THZ1 and LDC4297. Enhanced PIK3CA signaling leads to CDK7 phosphorylation, which promotes RNA Polymerase II phosphorylation and transcription. Milciclib, THZ1, and LDC4297 lead to a reduction in RNA Polymerase II phosphorylation on the SLC2A1 promoter. These data indicate that our high-throughput assay can identify compounds that regulate glucose consumption and that CDK7 is a key regulator of glucose consumption in cells with an activated PI3K pathway.